8.4 Van Der Riet 1

MICROALGAE FOR
AQUACULTURE:
CONVERTING ART
INTO TECHNOLOGY
ALGAEUROPE - Dec2016
TOMALGAE PRODUCES MICROALGAE
TOMALGAE PRODUCES MICROALGAE
FOR AQUACULTURE
“Algae are at the base of the entire aquatic food chain,…

Therefore, it is not surprising that the microalgae which compose the
phytoplankton play a vital role in the rearing of aquatic animals like
mollusks, shrimp, and fish, and have a strategic interest for aquaculture.”
(Arnaud Muller-Feuga 2000)

MICROALGAE IN AQUACULTURE
“Since artificial substitutes are inferior to live microalgae as feed for the
critical stages in the life cycles of several aqua-cultural species, a growing
demand for microalgae will go hand in hand with the expected growth of
aquaculture throughout the world.”
(Amos Richmond 2007)

MICROALGAE IN AQUACULTURE
• “Presently, most aquacultural enterprises produce (albeit with only limited

success in many cases) their own supply of microalgae.” (A. Richmond 2007)
• “Mass production of micro-algae has been recognized as a major bottle-neck

to many forms of marine hatchery and nursery production… The problem of
high costs of individual hatcheries producing their own algae is compounded
by the need of scarce expertise, without which crashes of algae at critical
periods occur quite commonly.” (Heasman et al. 2001)
• “This has prompted a search for alternatives to on-site production”
• The most obvious alternative: centralized microalgal facilities that produce,

concentrate and preserve the algae.
PRESERVED ALGAE FOR HATCHERIES:
OLD IDEA … NUMEROUS ATTEMPTS …
LITTLE SUCCESS
“Centralized microalgal facilities which sell (for a high price) frozen pastes or
highly concentrated refrigerated stock cultures cover at present only a small part
of the aquacultural demand for live microalgae.” (A. Richmond 2007)
And very often (typically)
“… commercial marine hatchery operators throughout the world have tried and
discarded substitutes for live micro-algae.” (Heasman 2001)
Obviously, further changes and innovative developments are required in this

field…
TOMALGAE’S FOCUS IS
THE DIATOMS (BACILLARIOPHYCEAE)
• The most productive and species-rich group of microalgae
• Are the foundation of ocean food web
• The most commonly grown microalgae on-site, specifically at shrimp and
shellfish hatcheries and nurseries.
Various
marine
planktonic
diatoms
TOMALGAE: SELECTING SUITABLE DIATOM
CULTIVAR
GENERAL ATTRIBUTES OF MICROALGAE USED IN AQUACULTURE:
- an appropriate size for ingestion

- readily digested
- rapid growth rates
- suitable for mass cultivation
- stable in culture
- good nutrient composition
- an absence of toxins
(M. R. Brown 2002)

FROM AN AQUACULTURE HATCHERY PERSPECTIVE, KEY
DESIRED ATTRIBUTES FOR MICROALGAL CONCENTRATES
ARE:
• High cell concentration without damage to cells

• Easy to suspend uniformly in water
• Neutrally or very slightly negatively buoyant cells
• Suitable nutritional composition
• Free from pathogens
• Appropriate packaging and shipping methods
• Acceptable shelf life
• Regularly available and affordable
R. Shields & I. Lupatsch, 2012
Heasman et al. 2001
10
DIATOMS USED IN AQUCULTURE
Chaetoceros spp.
11
DIATOMS USED IN AQUCULTURE
Thalassiosira spp.
12
DIATOMS USED IN AQUACULTURE
Cyclotella sp. Skeletonema sp.

DIATOMS USED IN AQUACULTURE
The diatoms (Bacillariophyta)
Phaeodactylum tricornutum
CHOICE OF TOMALGAE:
A ‘THALASSIOSIROID’ DIATOM
( ORDER THALASSIOSIRALES )
Thalassiosiroid diatoms grown commercially, closely related to the cultivar of

Tomalgae:
- Thalassiosira weissflogii
- Thalassiosira pseudonana
- Cyclotella cryptica
REED MARICULTURE INC
“Thalassiosira weissflogii … is used in the shrimp and shellfish larviculture

industry. This algae is considered by several hatcheries to be the single best
algae for larval shrimp.”
“Thalassiosira pseudonana is a small diatom … that is used in the shrimp

and bivalve larviculture industry. This alga has been found to be excellent for
larval shrimp.”
http://reedmariculture.com/product_instant_algae.php
SeaAg, Inc., founded 1989 Florida, Joe Weissman and
John Benemann, to produce littleneck clams on-shore
(grow diatoms in open ponds for seed clams and grow –out)
Cyclotella
grown in
open ponds, Clam hatchery -
fed to clams nursery
From
larvae
to seed
to
market
size
THALASSIOSIROID CULTIVAR OF TOMALGAE
COMMERCIAL NAME: THALAPURE
- Species-level taxonomy – required

revision.
- Cultivated in freshwater.
- Preservation – freeze-drying.
- Sufficient richness in PUFAs.
- Various workable applications,
but
current principal focus –
feeds for shrimp larvae
TOMALGAE: ACHIEVING ADEQUATE UNDERSTANDING
OF CULTIVAR BIOLOGY AND ITS INTERACTION WITH
ENVIRONMENT
Some important innovative developments and optimization of growth

conditions:
• Breeding program
• Temperature factor
• Rigorous control of culture contamination
• Nutritious quality control
• Securing the integrity of cells during concentration, preservation and
rehydration
DIATOMS: SEXUAL BREEDING
“In most diatom species, after one to several years

of size reduction, (but weeks in our intensive
culture) every surviving individual of every lineage
must reproduce sexually or die. Diatoms are some
of the most sexual organisms on earth;
our problem in understanding them is that we
do not invade their privacy often enough.”
Prof. Dr. D.G.Mann, 1999

20 BASICS ON THE LIFE CYCLE AND
SEXUAL BEHAVIOUR IN DIATOMS
Chepurnov V.A., et al. (2008). In

search of new tractable diatoms for
experimental biology. BioEssays 30:
692-702.
BREEDING PROGRAM FOR THE CULTIVAR
CURRENTLY FUTURE
Valvar cell size control: - Generating new strains and

Species-specific range: 3 - 45 µm; selection;
- Genetic control and
manipulations of economically
important traits;
- Domestication of the cultivar.
Required for early shrimp larvae
(zoea): ≤ 10 µm.
Control over triggering and intensity “Cross the best with the best,
of sexual reproduction select the best, and … hope for the best!”
(John W. Snape, 2004)
DR. JOHN BENEMANN (2010)
“We will have to domesticate algae, which has not yet

been done. We are still using wild types, that is, strains
that come directly out of nature without really any
significant improvement or even much selection.”
“… the strains are going to be eventually what drives

the industry, not somebody coming up with a better paddlewheel, or a more
clever way of injecting CO2, or some kind of fancy new photobioreactor.”
WHERE TO PRODUCE AT SCALE?
CLIMATIC OPTIMA FOR SHRIMP AND ALGAE CULTURE ARE
DIFFERENT
• Microalgae – temperature optimum: 18-23°C (Tomalgae cultivar: 15-20°C)

• Penaeid shrimps – temperature optimum: 29-33°C
• Thalapure – controlled greenhouse conditions, cultivated most of the year

at suitable temperature
• Thalapure – cultivated in freshwater environment (free of pathogens typical
for oceanic water)
RIGOROUS CONTROL OF CULTURE
CONTAMINATION
Grazers and parasites ‘Weed’ microalgae

PRESERVING THE QUALITY:
EFFICIENT DOWNSTREAM PROCESS
Pre-concentration Centrifugation
Freeze-dried microalgal powder
PRESERVING THE QUALITY: FREEZE DRYING & PACKAGING
THALAPURE:
AS FREEZE-DRIED POWDER
After freeze-drying,
nutritional quality of the powder remains
as high as the live algae, e.g. (PUFAs):
THALAPURE: KEEPING THE INTEGRITY OF
CELLS AT ALL STAGES OF GROWTH AND
PROCESSING
Concentrated biomass Dried biomass Rehydrated biomass

THALAPURE: AS FREEZE-DRIED POWDER
Correct rehydration
Incorrect rehydration
32
General health assessment of larval condition
Condition of the hepatopancreas and gut contents:
“In healthy larvae showing active feeding and digestion,

the hepatopancreas and midgut will be full of small,
easily observed bubbles (digestive or “lipid” vacuoles)…“
Health management and biosecurity maintenance in white shrimp (Penaeus

vannamei) hatcheries in Latin America, FAO Fisheries Technical Paper. No. 450. Rome, FAO. 2003.
62p. http://www.fao.org/docrep/007/y5040e/y5040e00.htm
33
PA C I F I C W H I T E S H R I M P : H E A LT H A S S E S S M E N T O F Z O E A
F E D W I T H T H A L A P U RE
hepatopancreas midgut
Zoea II
MICROALGAE FOR AQUACULTURE:
CONVERTING ART INTO TECHNOLOGY
Thalapure for shrimp hatcheries:
Converting ‘microalgae hatchery art’, i.e. diversity of farm- or company–

specific methods of on-site microalgae production
into technology, i.e.
Uniform, well-established and readily reproducible procedures.
The nearest realistic scenario: reducing dependence of hatcheries on live

microalgae especially at the most critical stages of larval rearing
TOMALGAE: MILESTONES
2013 (June - ) Pilot set-up at greenhouse location NL, focus on sustainable

production and quality. Test in target markets with selected
partner
2014 (Jan) Proof of concept first application (direct feed in larval AQ,
shrimp)
2014 (April) 1st commercial RW’s operational at site in Belgium
2014 (June) Supply agreement Thailand and China. First shipments
2014 (Dec) ISO-9001 and GMP+ certified
2015 (Feb) Tomalgae joined the Benchmark group
2015 Scale-up production facility (Phase 1)
(- present) • Upstream: 5000m2 (3750m2 net pond surface)
• Downstream: 2 x 20m3 xh-1 pre-concentration separation
technology
• Outsourcing: Freeze drying and packaging
BENCHMARK HOLDINGS — DIVISIONS
THANK YOU FOR YOUR
ATTENTION
William van der Riet &
Viktor Chepurnov

8.4 Van Der Riet 1

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MICROALGAE FOR

“Algae are at the base of the entire aquatic food chain,…

(Arnaud Muller-Feuga 2000)

(Amos Richmond 2007)

• “Presently, most aquacultural enterprises produce (albeit with only limited

• “Mass production of micro-algae has been recognized as a major bottle-neck

• “This has prompted a search for alternatives to on-site production”

• The most obvious alternative: centralized microalgal facilities that produce,

And very often (typically)

Obviously, further changes and innovative developments are required in this

- an appropriate size for ingestion

(M. R. Brown 2002)

• High cell concentration without damage to cells

Cyclotella sp. Skeletonema sp.

The diatoms (Bacillariophyta)

Thalassiosiroid diatoms grown commercially, closely related to the cultivar of

“Thalassiosira weissflogii … is used in the shrimp and shellfish larviculture

“Thalassiosira pseudonana is a small diatom … that is used in the shrimp

COMMERCIAL NAME: THALAPURE

- Species-level taxonomy – required

Some important innovative developments and optimization of growth

“In most diatom species, after one to several years

Prof. Dr. D.G.Mann, 1999

Chepurnov V.A., et al. (2008). In

Valvar cell size control: - Generating new strains and

“We will have to domesticate algae, which has not yet

“… the strains are going to be eventually what drives

• Microalgae – temperature optimum: 18-23°C (Tomalgae cultivar: 15-20°C)

• Thalapure – controlled greenhouse conditions, cultivated most of the year

Grazers and parasites ‘Weed’ microalgae

Concentrated biomass Dried biomass Rehydrated biomass

General health assessment of larval condition

Condition of the hepatopancreas and gut contents:

“In healthy larvae showing active feeding and digestion,

Health management and biosecurity maintenance in white shrimp (Penaeus

Converting ‘microalgae hatchery art’, i.e. diversity of farm- or company–

The nearest realistic scenario: reducing dependence of hatcheries on live

2013 (June - ) Pilot set-up at greenhouse location NL, focus on sustainable

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